DNA Structure and Function

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DNA Structure and Function

• Chapter 13

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Scientists that Studied DNA(know what they did
in general)

• Miescher
• Griffith
• Hershey and Chase
• Avery and Colleagues
• Pauling
• Chargaff
• Franklin
• Watson and Crick

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Miescher Discovered DNA

• 1868
• Johann Miescher investigated the chemical
  composition of the nucleus
• Isolated an organic acid that was high in
  phosphorus
• He called it nuclein
• We call it DNA (deoxyribonucleic acid)

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Mystery of the Hereditary Material

• Originally believed to be an unknown class of
  proteins
• Thinking was
• Heritable traits are diverse
• Molecules encoding traits must be diverse
• Proteins are made of 20 amino acids and are
  structurally diverse

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Griffith Discovers Transformation

• 1928
• Attempting to develop a vaccine
• Isolated two strains of Streptococcus pneumoniae
• Rough strain was harmless
• Smooth strain was pathogenic

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Griffith Discovers Transformation
1. Mice injected with live cells of harmless
strain R.
2. Mice injected with live cells of killer strain
S.
3. Mice injected with heat-killed S cells.
4. Mice injected with live R cells plus
heat-killed S cells.
Mice die. Live S cells in their blood.
Mice live. No live R cells in their blood.
Mice die. Live S cells in their blood.
Mice live. No live S cells in their blood.
Figure 13.3Page 218
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Transformation

• What happened in the fourth experiment?
• The harmless R cells had been transformed by
  material from the dead S cells
• Descendents of the transformed cells were also
  pathogenic

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Bacteriophages
Protein coat

• Viruses that infect bacteria
• Consist of protein and DNA
• Inject their hereditary material into bacteria

bacterial cell wall
plasma membrane
cytoplasm
DNA
Figure 13.4bPage 219
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Hershey Chases Experiments (1940s)

• Created labeled bacteriophages
• Radioactive sulfur
• Radioactive phosphorus
• Allowed labeled viruses to infect bacteria
• Asked Where are the radioactive labels after
  infection?

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Hershey and Chase Results
virus particle labeled with 35S
virus particle labeled with 32P
bacterial cell (cutaway view)
label outside cell
label inside cell
Figure 13.5Page 219
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Avery and Colleagues (1944)

• What is the transforming material?
• Cell extracts treated with protein-digesting
  enzymes could still transform bacteria
• Cell extracts treated with DNA-digesting enzymes
  lost their transforming ability
• Concluded that DNA, not protein, transforms
  bacteria TRANSFORMING PRINCIPLE

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Structure of the Hereditary Material

• Experiments in the 1950s showed that DNA is the
  hereditary material
• Scientists raced to determine the structure of
  DNA
• 1951 Linus Pauling discovered the 3D structure
  of the protein collagen

Figure 13.2Page 217
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Structure of Nucleotides in DNA

• Each nucleotide consists of
• Deoxyribose (5-carbon sugar aka pentose)
• Phosphate group
• A nitrogen-containing base
• Four bases
• Adenine, Guanine, Thymine, Cytosine

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Nucleotide Bases
ADENINE (A) PURINE
GUANINE (G) PURINE
phosphate group
deoxyribose
THYMINE (T) PYRIMIDINE
CYTOSINE (C) PYRIMIDINE
Figure 13.6Page 220
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Animation
DNA subunits interaction.
Click to view animation.
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Composition of DNA

• Chargaff showed
• Amount of adenine relative to guanine differs
  among species
• Amount of adenine always equals amount of thymine
  and amount of guanine always equals amount of
  cytosine
• AT and GC

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Rosalind Franklins Work

• Was an expert in X-ray crystallography
• Used this technique to examine DNA fibers
• Concluded that DNA was some sort of helix

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Watson-Crick Model

• DNA consists of two nucleotide strands
• Strands run in opposite directions (5 prime to 3
  prime and 3 to 5)
• Strands are held together by hydrogen bonds
  between bases
• A binds with T and C with G
• Molecule is a double helix

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Watson-Crick Model
Figure 13.7Page 221
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Animation
DNA close-up animation.
Click to view animation.
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Animation
DNA structure animation.
Click to view animation.
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DNA Structure Helps Explain How It Duplicates

• DNA is two nucleotide strands held together by
  hydrogen bonds
• Hydrogen bonds between two strands are easily
  broken
• Each single strand then serves as template for
  new strand

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DNA Replication

• Each parent strand remains intact
• Every DNA molecule is half old and half new
  semi-conservative
• 1 strand of the original is 25 of the
  transcribed helix

new
new
old
old
Figure 13.9Page 222
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Base Pairing during Replication

• Each old strand serves as the template for
  complementary new strand

Figure 13.10Page 223
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Enzymes in Replication

• DNA polymerase
• Unwind the strands
• attaches complementary nucleotides (replication
  enzyme)
• Proof reads replicated strands
• Repairs DNA
• DNA ligase
• fills in gaps of nucleotides into one continuous
  strand
• Repairs DNA
• Wind the two strands together

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Replication (hereditary material duplicated)
happens prior to cell division (mitosis)
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A Closer Look at Strand Assembly

• Energy for strand assembly is provided by
  removal of two phosphate groups from free
  nucleotides

newly forming DNA strand
one parent DNA strand
Figure 13.10Page 223
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Continuous and Discontinuous Assembly
3
5
Strands can only be assembled in the 5 to 3
direction
Discontinuous
Continuous 5
3
Figure 13.10Page 223
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Animation
DNA replication animation.
Click to view animation.
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DNA Repair

• Mistakes can occur during replication
• DNA polymerase can
• Proof read sequence from complementary strand
• Fixes mistakes in strand with DNA ligase
• Genetic disorders if DNA repair skipping the
  wrong sequences (mutations)

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Cloning

• Making a genetically identical copy of an
  individual
• Researchers have been creating clones for decades
• These clones were created by embryo splitting

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Dolly Cloned from an Adult Cell

• Showed that differentiated cells could be used to
  create clones
• Sheep udder cell was combined with enucleated
  (nucleus taken out) egg cell
• Dolly is genetically identical to the sheep that
  donated the udder cell

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Animation
Cloning of Dolly animation.
Click to view animation.
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More Clones

• Mice
• Cows
• Pigs
• Goats
• Guar (endangered species)